EZ-Tn5™ Tnp Transposome™ Kits < DHFR-1> or < KAN-2>

Catalogue number

Pack size

TSM99D1

10 reactions <DHFR-1>

TSM99K2

10 reactions <KAN-2>

EZ-Tn5™ Transposase

Catalogue number

Pack size

TNP92110

10 units

Sterile Nuclease-free Water

Catalogue number

Pack size

W7350ML

50ml

EZ-Tn5™ Transposome™ complexs are formed between an EZ-Tn5™ Transposon and EZ-Tn5™ Transposase and provide a simple and reliable method for generating a library of random gene knockouts in vivo. Just electroporate the EZ-Tn5™ Transposome into any of a broad range of living cells and select for a marker encoded by the EZ-Tn5™ Transposon (Figure 1). Because there is no need for cell conjugation, suicide vectors, or specific host factors, EZ-Tn5™ Transposomes are ideal for creating mutants in species that have poorly described genetic systems or lack adequate molecular tools.

Ready-to-use EZ-Tn5™ Transposomes are available containing either a kanamycin selectable marker (<KAN-2>) or dihydrofolate reductase gene (<DHFR-1>) that can be selected on plates containing trimethoprim. These markers are readily expressed in many gram-negative bacteria. Or you can create your own EZ-Tn5™ Transposome using one of the EZ-Tn5™ pMOD™ Transposon Construction Vectors and EZ-Tn5™ Transposase.

All EZ-Tn5™ Transposons contain unique primer binding sites at each end for bidirectional sequencing. Hence, a gene knockout can be sequenced directly using bacterial genomic DNA as template and the primers provided with each Transposome. Transposon insertions made using an EZ-Tn5™ <R6Kγori/KAN-2>Tnp Transposome™ Kit or HyperMu™ lt;R6Kγori/KAN-1> Tnp Transposome™ Kit with a transposon can be rescued and the flanking DNA sequenced.

EZ-Tn5™ Transposome-mediated insertions have been made in many different microorganisms, including Acinetobactor, Campylobacter, Escherichia, Mycobacterium, Proteus, Pseudomonas, Saccharomyces, Salmonella, Trypanosoma, Xylella, and others. The number of transposition clones obtained is highly dependent on the transformation efficiency of the host cel.

The presence of one or more type I restriction and modification systems may lower transposition efficiencies. Epicentre® scientists have shown that these systems can be blocked in vivo with the TypeOne™ Restriction Inhibitor.

Figure 1. The EZ-Tn5™ Transposon insertion site in bacterial DNA can be sequenced directly using genomic DNA isolated using the MasterPure™ DNA Purification Kit and primers homologous to the ends of the transposon.

Last modified: 2008-05-14 16:46:46

Protocols for: EZ-Tn5™ Transposome™ Kits

EZ-Tn5™ <KAN-2> Transposome™ Kit Protocol

(catalogue number TSM99K2)

EZ-Tn5™ <KAN-2> Transposon / KAN-2 FP-1 Forward Primer / KAN-2 RP-1 Reverse Primer Protocol

(catalogue number KAN2001 / KFP9122 / KRP0021)

EZ-Tn5™ <DHFR-1> Transposome™ Kit Protocol

(catalogue number TSM99D1)

EZ-Tn5™ <DHFR-1> Transposon / DHFR-1 FP-1 Forward Primer / DHRP-1 RP-1Reverse Primer Protocol

(catalogue number DFR1001 / DFP9121 / DRP9131)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46